LOCUS       sequence-252560-        4263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Phytopathogen Pseudomonas syringae pv. tomato type III secretion 
            system effector (gene lacking stop codon) Gateway donor clone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4263)
  AUTHORS   Munkvold KR, Martin ME, Bronstein PA, Collmer A
  TITLE     A survey of the Pseudomonas syringae pv. tomato DC3000 type III 
            secretion system effector repertoire reveals several effectors that 
            are deleterious when expressed in Saccharomyces cerevisiae.
  JOURNAL   Mol Plant Microbe Interact. 2008 Apr;21(4):490-502. doi: 
            10.1094/MPMI-21-4-0490.
  PUBMED    18321194
REFERENCE   2  (bases 1 to 4263)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Mol Plant Microbe Interact"; date: "2008-04"; volume: "21"; issue: 
            "4"; pages: "490-502"
COMMENT     Sequence Label: pCPP5616
FEATURES             Location/Qualifiers
     source          1..4263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1688..1787)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1804..1823)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1805..1823)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1828..1844)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1828..1844)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1946..1965)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1957..2766
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2024..2043)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2913..3501
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3402..3421
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3655..3672
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3831..3858)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3950..4036)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4085..4107
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4099..4116
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4100..4116
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4132..4231
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     RBS             4243..4263
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 cccttcacca tggcgggtat caatagagcg ggaccatcgg gcgcttattt tgttggccac
       61 acagaccccg agccagtatc ggggcaagca cacggatccg gcagcggcgc cagctcctcg
      121 aacagtccgc aggttcagcc gcgaccctcg aatactcccc cgtcgaacgc gcccgcaccg
      181 ccgccaaccg gacgtgagag gctttcacga tccacggcgc tgtcgcgcca aaccagggag
      241 tggctggagc agggtatgcc tacagcggag gatgccagcg tgcgtcgtag gccacaggtg
      301 actgccgatg ccgcaacgcc gcgtgcagag gcaagacgca cgccggaggc aactgccgat
      361 gccagcgcac cgcgtagagg ggcggttgca cacgccaaca gtatcgttca gcaattggtc
      421 agtgagggcg ctgatatttc gcatactcgt aacatgctcc gcaatgcaat gaatggcgac
      481 gcagtcgctt tttctcgagt agaacagaac atatttcgcc agcatttccc gaacatgccc
      541 atgcatggaa tcagccgaga ttcggaactc gctatcgagc tccgtggggc gcttcgtcga
      601 gcggttcacc aacaggcggc gtcagcgcca gtgaggtcgc ccacgccaac accggccagc
      661 cctgcggcat catcatcggg cagcagtcag cgttctttat ttggacggtt tgcccgtttg
      721 atggcgccaa accagggacg gtcgtcgaac actgccgcct ctcagacgcc ggtcgacagg
      781 agcccgccac gcgtcaacca aagacccata cgcgtcgaca gggctgcgat gcgtaatcgt
      841 ggcaatgacg aggcggacgc cgcgctgcgg gggttagtac aacagggggt caatttagag
      901 cacctgcgca cggcccttga aagacatgta atgcagcgcc tccctatccc cctcgatata
      961 ggcagcgcgt tgcagaatgt gggaattaac ccaagtatcg acttggggga aagccttgtg
     1021 caacatcccc tgctgaattt gaatgtagcg ttgaatcgca tgctggggct gcgtcccagc
     1081 gctgaaagag cgcctcgtcc agccgtcccc gtggctcccg cgaccgcctc caggcgaccg
     1141 gatggtacgc gtgcaacacg attgcgggtg atgccggagc gggaggatta cgaaaataat
     1201 gtggcttatg gagtgcgctt gcttaacctg aacccggggg tgggggtaag gcaggctgtt
     1261 gcggcctttg taaccgaccg ggctgagcgg ccagcagtgg tggctaatat ccgggcagcc
     1321 ctggacccta tcgcgtcaca attcagtcag ctgcgcacaa tttcgaaggc cgatgctgaa
     1381 tctgaagagc tgggttttaa ggatgcggca gatcatcaca cggatgacgt gacgcactgt
     1441 ctttttggcg gagaattgtc gctgagtaat ccggatcagc aggtgatcgg tttggcgggt
     1501 aatccgacgg acacgtcgca gccttacagc caagagggaa ataaggacct ggcgttcatg
     1561 gatatgaaaa aacttgccca attcctcgca ggcaagcctg agcatccgat gaccagagaa
     1621 acgcttaacg ccgaaaatat cgccaagtat gcttttagaa tagtcccctg caagggtggg
     1681 cgcgccgacc cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt
     1741 tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat
     1801 atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg
     1861 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa
     1921 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
     1981 tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     2041 gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca
     2101 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     2161 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     2221 cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta
     2281 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     2341 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     2401 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     2461 aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg
     2521 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     2581 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     2641 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     2701 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     2761 ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg
     2821 gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg
     2881 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2941 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3001 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     3061 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3121 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3181 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3241 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3301 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3361 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3421 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3481 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3541 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3601 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     3661 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     3721 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     3781 aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat
     3841 ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc
     3901 gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac
     3961 tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag
     4021 cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg
     4081 ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc ccaaataatg
     4141 attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgatgagca atgctttttt
     4201 ataatgccaa ctttgtacaa aaaagcaggc tccgcggccg ccttgtttaa ctttaagaag
     4261 gag
//
